{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T09:46:01Z","timestamp":1791107161325,"version":"4.1.0"},"reference-count":50,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2003,9,3]],"date-time":"2003-09-03T00:00:00Z","timestamp":1062547200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microscopy Res &amp; Technique"],"published-print":{"date-parts":[[2003,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Though molecular biology\u2010based visualization techniques such as antibody staining, in situ hybridization, and induction of reporter gene expression have become routine procedures for analyzing the structures of the brain, precautions to prevent misinterpretation have not always been taken when preparing and interpreting images. For example, sigmoidal development of the chemical processes in staining might exaggerate the specificity of a label. Or, adjustment of exposure for bright fluorescent signals might result in overlooking weak signals. Furthermore, documentation of a staining pattern is affected easily by recognized organized features in the image while other parts interpreted as \u201cdisorganized\u201d may be ignored or discounted. Also, a higher intensity of a label per cell can often be confused with a higher percentage of labeled cells among a population. The quality, and hence interpretability, of the three\u2010dimensional reconstruction with confocal microscopy can be affected by the attenuation of fluorescence during the scan, the refraction between the immersion and mounting media, and the choice of the reconstruction algorithm. Additionally, visualization of neurons with the induced expression of reporter genes can suffer because of the low specificity and low ubiquity of the expression drivers. The morphology and even the number of labeled cells can differ considerably depending on the reporters and antibodies used for detection. These aspects might affect the reliability of the experiments that involves induced expression of effector genes to perturb cellular functions. Examples of these potential pitfalls are discussed here using staining of <jats:italic>Drosophila<\/jats:italic> brain. Microsc. Res. Tech. 62:170\u2013186, 2003. \u00a9 2003 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/jemt.10369","type":"journal-article","created":{"date-parts":[[2003,9,5]],"date-time":"2003-09-05T10:44:19Z","timestamp":1062758659000},"page":"170-186","source":"Crossref","is-referenced-by-count":47,"title":["Cautionary observations on preparing and interpreting brain images using molecular biology\u2010based staining techniques"],"prefix":"10.1002","volume":"62","author":[{"given":"Kei","family":"Ito","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryuichi","family":"Okada","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nobuaki K.","family":"Tanaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takeshi","family":"Awasaki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2003,9,3]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.97.22.11984"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1177\/002215549804600911"},{"key":"e_1_2_1_4_1","series-title":"Experimental neuroanatomy: the practical approach series","first-page":"273","author":"Bolam JP","year":"1992"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/0959-4388(95)80061-1"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.118.2.401"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.13.5972"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.76.5.2192-2198.2002"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0596-606"},{"key":"e_1_2_1_10_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.274.5295.2104"},{"key":"e_1_2_1_11_1","doi-asserted-by":"publisher","DOI":"10.1006\/dbio.1995.1115"},{"key":"e_1_2_1_12_1","doi-asserted-by":"publisher","DOI":"10.3109\/01677060009084496"},{"key":"e_1_2_1_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)00084-2"},{"key":"e_1_2_1_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0960-9822(02)01239-3"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/332853a0"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00636536"},{"key":"e_1_2_1_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(91)90571-F"},{"key":"e_1_2_1_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/gene.10137"},{"key":"e_1_2_1_19_1","first-page":"679","volume-title":"Selected methods for antibody and nucleic acid probes","author":"Hockfield S","year":"1993"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF02179499"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.124.4.761"},{"key":"e_1_2_1_22_1","doi-asserted-by":"publisher","DOI":"10.1007\/s004410050901"},{"key":"e_1_2_1_23_1","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1101\/lm.5.1.52","article-title":"The organization of extrinsic neurons and their implications regarding the functional roles of the mushroom bodies in Drosophila melanogaster Meigen","volume":"5","author":"Ito K","year":"1998","journal-title":"Learn Mem"},{"key":"e_1_2_1_24_1","doi-asserted-by":"publisher","DOI":"10.1002\/bies.950131206"},{"key":"e_1_2_1_25_1","doi-asserted-by":"publisher","DOI":"10.1002\/neu.10100"},{"key":"e_1_2_1_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)80701-1"},{"key":"e_1_2_1_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0091-679X(08)60933-4"},{"key":"e_1_2_1_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/0896-6273(94)90022-1"},{"key":"e_1_2_1_29_1","doi-asserted-by":"publisher","DOI":"10.3109\/01677060109167373"},{"key":"e_1_2_1_30_1","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1093\/genetics\/157.2.727","article-title":"Dual\u2010tagging gene trap of novel genes in Drosophila melanogaster","volume":"157","author":"Lukacsovich T","year":"2001","journal-title":"Genetics"},{"key":"e_1_2_1_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(02)00700-6"},{"key":"e_1_2_1_32_1","doi-asserted-by":"publisher","DOI":"10.2144\/99265rr01"},{"key":"e_1_2_1_33_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-4695(199812)37:4<607::AID-NEU9>3.0.CO;2-Q"},{"key":"e_1_2_1_34_1","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1998.2075"},{"key":"e_1_2_1_35_1","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1093\/oso\/9780199636617.003.0005","volume-title":"Drosophila, a practical approach","author":"O'Kane CJ","year":"1998"},{"key":"e_1_2_1_36_1","doi-asserted-by":"crossref","unstructured":"O'KaneCJ GehringWJ.1987.Detection in situ of genomic regulatory elements inDrosophila. 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